Potential sources of microbiological spoilage in beer brewing
Potential sources of primary contamination
Brewing water: e.g. Enterobacteriaceae
Yeast slurry: contamination with e.g. Lactobacillus, non cultured yeast as e.g. S. diastaticus
optional aroma hops
The GEN-IAL kits enable the detection and identification of beer spoilage bacteria and yeast in various sample types including in-process and final product samples.
Potential sources of secondary contamination
After filtration: main sources for detection of different beer spoilers
Filling station: e.g. S. diastaticus
Product: final quality product control, bacteria /yeast
malt
bitter hops
Faster and more sensitive results with real-time PCR – customize your routine analysis
Either extremely fast or maximal sensitive detection or specific identification is possible by combining different methods of sample preparation and detection with real-time PCR.
Approaches for microbiological analysis in beer
Time requirement: ~ 2 hrs
Centrifugation and qPCR
• For in-process controls (yeast-containing samples and filtrate)
• Kit for detection of bacteria and (wild) yeast in yeast containing samples (yeast propagation or fermentation tank)
• Extremely fast
• Screening and/or identification
• Only two hours to result
Time requirement: ~ 2 hrs
Filtration and qPCR
• Fast screening approach for specific applications
• Volume maximum 30 mL
• Sensitivity 10 - 100 cfu/30 mL (without enrichment)
• High sensitivity with enrichment
• Extremely fast
• Screening and/or identification
• Only two hours to result
Time requirement: ~ 50 hrs
Enrichment and qPCR
• Volume maximum 1 L (depending on sample type)
• Sensitivity approx. 100 cfus
• Pre-enrichment and qPCR: minimum 2 days and 2 hours from sample to result
• Modern established approach for beverage analytic
• Specific flow chart documents for different applications are available (e.g. for filtration and incubation or filtration and direct detection)
• Fast
• Screening and/or identification
• Ensure abscence of low cfu/sample volume
Precoated PCR strips – a unique solution for convenient handling
Different combinations of screening and identification possibilities are available, thus allowing a cost effective and customized routine analysis.
The QuickGEN kits contain 8-well strips which are precoated with the reagents for up to 4 different parameters per tube. Each tube of a 8-well strip may contain different parameters: this allows a multiplex panel for up to 4 x 8 parameters for one sample in a 8-well strip.
Templates are available for MyGo Pro and BioRad CFX 96. These prepared templates contain the settings for dedicated kits and allow a direct start of the real-time PCR without the need to program the settings:
Start run 1 2 3
Open the template of dedicated kit/parameter
Add the sample names
• Agilent MX 3005P*
• ABI 7500 or higher*
• Thermo Fischer QuantStudio™ 5 or higher (0.2 ml block)
• Bioer QuantGene 9600
• MyGo Pro or ESR */**
• Thermo Fischer QuantStudio™ 5 or higher (0.1 ml block)
• Roche Lightcycler® 480 II
• BioRad® Opus CFX 96
• BioRad® CFX 96
• Analytik Jena qTOWER / iris
* discontinued but still on the market
** 4plex assays for MyGo Pro and ESR requires a specific kit (Atto490 instead of Cy5) with number Qxy4 (e.g. Q024, Q044)
1. Group specific screening
An initial screening gives a group specific answer of for example yeast and bacteria in one assay. Different group specific screening combinations are available:
3plex Lactobacillus/Pediococcus, Megasphaera/Pectinatus, IAC – Q03z kits
4plex Lactobacillus/Pediococcus, Megasphaera/Pectinatus, IAC
IAC: Internal Amplification Control or Internal Control
2. Screening and differentiation in one assay
Yeast
S. disastacicus Dekkera bruxellensis Q02z kits
Q04z kits Q09z kits
The most relevant beer spoilage bacteria and yeast can be identified in one assay for example with kit Q081 - Q083. 12 strips of 8 wells allows for the detection of the most relevant spoilage organisms for 12 samples in one assay.
1 NTC NTC IAC
2 Enterobacteriaceae P. anomala Saccharomyces cerevisae var. diastaticus
3 P. damnosus P. acidilactici/pentosaceus/parvulus/inopinatus P. claussenii
4 Pectinatus spp. Megasphaera spp. L. rossiae
5 L. brevis/L.parabrevis/L.brevisimilis L. lindneri L. casei/L.paracasei
6 L. buchneri/L. parabuchneri L. collinoides/L. paracollinoides L. perolens/L. harbinensis
7 L. plantarum/L. paraplantarum L. coryniformis IAC
8 L. acetotolerans L. backii PTC
High Low White
Qxy1 Qxy2 Qxy3
Detection of bacteria and yeasts: the kit Q571-Q573 allows the detection of bacteria and wild yeast screening in yeast containing samples
Tube Sample FAM HEX ROX
1 NTC – – IAC
2 PTC Positive Control – –
3 1 Lactobacillus/ Pediococcus Megasphaera/ Pectinatus Acetic acid bacteria
4 1 – Wild yeast 1* IAC
5 1 – Wild yeast 2* –
6 2 Lactobacillus/ Pediococcus Megasphaera/ Pectinatus Acetic acid bacteria
7 2 – Wild yeast 1* IAC
8 2 – Wild yeast 2* –
3. Detection and identification of yeast or bacteria only
Detection of bacteria and yeasts: the kit Q071-Q073 allows the detection of bacteria and wild yeast screening for filtered samples
Tube Sample FAM HEX ROX
1 NTC – – IAC
2 PTC Positive Control – –
3 1 Enterobacteriaceae Lactobacillus/ Pediococcus Pediococcus
4 1 Wild yeast 1* Bottom fermented yeast IAC
5 1 Wild yeast 2* Top fermented yeast Acetic acid bacteria
6 2 Enterobacteriaceae Lactobacillus/ Pediococcus Pediococcus
7 2 Wild yeast 1* Bottom fermented yeast IAC
8 2 Wild yeast 2* Top fermented yeast Acetic acid bacteria
Several kits are available for the detection of specific bacteria or yeast only. The kit Q541-Q543 allow the identification of 12 yeast species per sample. 12 samples can be tested per kit (12 strips).
Tube FAM HEX
1 NTC IAC
2 Rhodotorula spp. Saccharomyces exiguus
3 Candida spp. Saccharomyces cerevisae var. diastaticus
4 Saccharomycodes ludwigii Debaromyces hansenii
5 Torulaspora delbrückii Saccharomyces bayanus/pastorianus
6 Kluyveromyces marxianus Hanseniaspora spp.
7 Dekkera spp. IAC
8 Pichia spp. PTC
* Wild yeast 1: Dekkera anomala, Dekkera bruxellensis, Dekkera custersiana, Dekkera naardenensis, Debaromyces hansenii, Hanseniaspora guillermondii, Hanseniaspora osmophila, Hanseniaspora uvarum, Issotchenkia orientalis, Kazachstania Exigua, Kluyveromyces marxianus, Metschnikowia pulcherrina, Pichia anomala, Pichia fermentans, Pichia membranaefaciens, Saccharomyces cerevisiae var. diastaticus, Saccharomycodes ludwigii, Torulaspora delbrückii
* Wild yeast 2: Candida glabrata, Candida albicans, Candida kefyr, Candida intermedia, Candida parapsilosis, Candida sake, Candida tropicalis, Naumovozyma dairenensis, Pichia guilliermondii, Zygosaccharomyces bailii, Zygosaccharomyces rouxii
The kit Q201-203 allows the identification and differentiation of bacteria that are usually classified as beer spoilage bacteria. Other relevant but often not directly as spoilage classified Lactobacillus bacteria are not included in this screening.
Tube Sample FAM HEX ROX
1 NTC Negative Control – IAC
2 PTC Positive Control – –
3 1 L. rossiae L. backii IAC
4 1 L. brevis/ parabrevis/ brevismillis L. lindneri L. casei/paracasei
5 1 Pediococcus spp. Lactobacillus spp. Megasphaera/ Pectinatus
6 2 L. rossiae L. backii IAC
7 2 L. brevis/ parabrevis/ brevismillis L. lindneri L. casei/paracasei
8 2 Pediococcus spp. Lactobacillus spp. Megasphaera/ Pectinatus
* Pediococcus spp.: P. damnosus, P. claussenii, P. acidilactici, P. pentosaceus, P. parvulus, P. inopinatus
* Lactobacillus spp.: L. collinoides / paracollinoides, L. coryniformis, L. plantarum/paraplantarum, L. perplens/harbinesis, L. acetolarans, L. buchneri/parabuchneri
GEN-IAL® – products for beer analysis
Product Description
Beer DNA preparation
GEN-IAL®Simplex® Easy DNA kit * DNA preparation of beverage samples
GEN-IAL® QuickGEN Sample preparation filtration DNA preparation of beverage samples, filtration
GEN-IAL® QuickGEN Sample preparation centrifugation
DNA preparation of beverage samples, centrifugation
GEN-IAL® QuickGEN Sample preparation in yeast For beverage samples mainly containing yeast
Automatic Magnetic DNA Extraction Kit
Beer – bacteria & yeast
GEN-IAL® QuickGEN First-Beer Differentiation PCR Kit
GEN-IAL® QuickGEN First-Beer Differentiation PCR Kit Screen, identification of spoilage bacteria
GEN-IAL® QuickGEN First-Beer yeast and bacteria differentiation
GEN-IAL® QuickGEN First-Beer yeast and bacteria differentiation for yeast containing samples
GEN-IAL® QuickGEN P1 Screening of bacteria and yeast
GEN-IAL® QuickGEN P1 Screening of bacteria and yeast
GEN-IAL® QuickGEN P1 Screening of bacteria and S. diastaticus
GEN-IAL® QuickGEN P1 Screening of bacteria and S. diastaticus
GEN-IAL® QuickGEN P1 Screening of bacteria and Dekkera spp.
GEN-IAL® QuickGEN* First-Biofilm
Beer – bacteria
GEN-IAL® QuickGEN P1 Screening of bacteria without yeast
GEN-IAL® QuickGEN P1 Screening of bacteria without yeast
GEN-IAL® QuickGEN P1 Screening of bacteria and Hop resistance
GEN-IAL® QuickGEN Pectinatus spp./Megasphaera spp.
GEN-IAL® QuickGEN Pectinatus spp./Megasphaera spp.
GEN-IAL® QuickGEN Enterobacteriaceae
GEN-IAL® QuickGEN Acetic acid bacteria
Automated extraction of bacteria- and yeast-DNA
Qualitative multiplex real-time PCR
Multiplex detection (30 species) and identification (19 species) of relevant beer spoilers See table page 4/5
Multiplex detection of classified relevant beer spoilage bacteria See table page 4/5
Multiplex detection and identification of beverage spoiling bacteria and yeasts See table page 4/5
Multiplex detection and identification of beverage spoiling bacteria and yeasts See table, page 4/5
DNA screening and differentiation of beer spoiling bacteria and yeasts (Lactobacillus, Pediococcus/Megasphaera, Pectinatus)
DNA screening and differentiation of beer spoiling bacteria and yeasts (Lactobacillus, Pediococcus/Megasphaera, Pectinatus/yeast)
DNA screening and differentiation of beer spoiling bacteria and Saccharomyces cerevisiae var. diastaticus
DNA screening and differentiation of beer spoiling bacteria and Saccharomyces cerevisiae var. diastaticus
DNA screening and differentiation of beer spoiling bacteria and yeasts (Lactobacillus, Pediococcus/Megasphaera, Pectinatus/ Dekkera spp.)
DNA detection of Lactococcus lactis, Leuconostoc mesenteroides and Pichia anomala
Qualitative real-time PCR
DNA screening and differentiation of beer spoiling bacteria (Lactobacillus, Pediococcus/Megasphaera, Pectinatus)
DNA screening of yeast slurry samples
DNA screening and differentiation of beer spoiling bacteria and hop resistance genes horA/horC
DNA detection and differentiation of Pectinatus and Megasphaera
DNA detection and differentiation of Pectinatus and Megasphaera
DNA detection of Enterobacteriaceae
DNA detection of acetic acid bacteria
100 preparations Q001
100 preparations Q004
100 preparations Q002
100 preparations Q005
50 preparations Q007
96 reactions/12 samples Q081 Q082 Q083
No. of tests Art. No. New
96 reactions/24 samples Q201 Q202 Q203
96 reactions/24 samples Q071 Q072 Q073
96/reactions/24 samples Q571 Q572 Q573
48 reactions Q021 Q022 Q023 Q024
50 reactions Q025
48 reactions Q041 Q042 Q043 Q044
50 reactions Q045
48 reactions Q091 Q092 Q093 Q094
50 reactions Q095
48 reactions Q031 Q032 Q033
48 reactions Q034
48 reactions Q051 Q052 Q053 Q054
50 reactions Q927
48 reactions Q112
50 reactions Q145
48 reactions Q511 Q512 Q513 Q514
Product Description
Beer – resistance genes
GEN-IAL®QuickGEN hop resistance genes hor A and hor C/hit A and orf5
Beer – yeast
GEN-IAL® QuickGEN First-Yeast PCR Kit Wild Yeast 1
GEN-IAL® QuickGEN Yeast PCR Kit
GEN-IAL® QuickGEN First-Yeast PCR Kit Wild Yeast 2
GEN-IAL® QuickGEN First-Yeast differentiation PCR Kit
GEN-IAL® QuickGEN® Yeast Dekkera bruxellensis
GEN-IAL® Pichia anomala*
GEN-IAL® Saccharomyces diastaticus*
GEN-IAL® Bottom fermented yeast*
GEN-IAL® QuickGEN Bottom fermented yeast
GEN-IAL® Top fermented yeast*
GEN-IAL® QuickGEN Top fermented yeast
GEN-IAL® QuickGEN Yeast Zygosachharomyces bailii
Qualitative real-time PCR
DNA detection of hop resistance genes
Qualitative real-time PCR
DNA screening and differentiation of wild yeast 1
DNA Detection of detection of S. cerevisiae var. diastaticus and Dekkera spp. in beverages
DNA screening and differentiation of wild yeast 2
DNA screening and differentiation of 12 yeasts
DNA detection of Dekkera bruxellensis
DNA detection of Pichia anomala (Wickerhamomyces anomalus)
DNA detection of Saccharomyces cerevisiae var. diastaticus
DNA detection of bottom fermented yeast
DNA detection of bottom fermented yeast
DNA detection of top fermented yeast
DNA detection of top fermented yeast
DNA detection of Zygosaccharomyces bailii
GEN-IAL® accessories Real-time PCR
GEN-IAL® Dekkera bruxellensis Standards
Color Compensation Kit LightCycler® 480
Washing solution
DNA standards for Dekkera bruxellensis quantification
Color compensation kit for multiplex assays
Washing solution for Q300
50 reactions Q105
50 reactions Q525
48 reactions Q213
50 reactions Q535
96 reactions/12 samples Q541 Q542 Q543
48 reactions Q371 Q372 Q373
50 reactions Q175
50 reactions Q934
50 reactions Q933
48 reactions Q161 Q162 Q163
50 reactions Q931
48 reactions Q151 Q152 Q153
48 reactions Q561 Q562 Q563
200.000 cfu Q360
5 reactions Q800
43 ml Q301
Wild yeast 1: Dekkera anomala, Dekkera bruxellensis, Dekkera custersiana, Dekkera naardenensis, Debaromyces hansenii, Hanseniaspora guillermondii, Hanseniaspora osmophila, Hanseniaspora uvarum, Issotchenkia orientalis, Kazachstania Exigua, Kluyveromyces marxianus, Metschnikowia pulcherrina, Pichia anomala, Pichia fermentans, Pichia membranaefaciens, Saccharomyces cerevisiae var. diastaticus, Saccharomycodes ludwigii, Torulaspora delbrückii
Wild yeast 2: Candida glabrata, Candida albicans, Candida kefyr, Candida intermedia, Candida parapsilosis, Candida sake, Candida tropicalis, Naumovozyma dairenensis, Pichia guilliermondii, Zygosaccharomyces bailii, Zygosaccharomyces rouxii
* Please be aware, for non-QuickGEN detection kits, the DNA preparation kit Q001 must be used, not Q002-Q005.
Benefits
+ DNA Extraction – easy one-step DNA lysis method
• One DNA extraction method for the detection of bacteria and yeast
• The yeast lysis is integrated in the qPCR reaction - no external mechanical disruptor, ultrasonic bath or heating block is required.
• Reduced handling errors
+ Real-time PCR assays
• Ready to use consumables – tube stripes pre-coated with lyticase, primer and probes (depending on the thermocycler, different versions of kits are available, further information is available on request)
• Simultaneous detection of the most relevant beer spoiling bacteria and yeast in one assay
Temperature Profile
Contact your R-Biopharm sales representative for more information.